Rho激酶抑制剂法舒地尔通过抑制MLK3－JNK信号通路保护脑缺血再灌注大鼠海马CAl区神经元被引量：1

Rho kinase inhibitor fasudil protect neurons in hippocampal CA1 region following cerebral ischenda reperfusion through inhibiting MLK3-JNK signal transduction pathway in rats

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摘要目的探讨Rho激酶抑制剂法舒地尔对脑缺血再灌注大鼠混合性谱系激酶（mixed—lineagekinase3，MLK3）、c—Jun—N末端激酶（c-JunNH2-terminalkinase，JNK）磷酸化、胱冬酶-3表达以及海马CAl区神经元损伤的影响。方法72只Sprague—Dawley大鼠随机分为假手术组、缺血再灌注组、生理盐水对照组和法舒地尔组。四血管结扎法制作大鼠全脑缺血模型。免疫印迹分析检测MLK3和JNK磷酸化水平以及胱冬酶-3表达，采用焦油紫染色图像分析检测海马CAl区存活神经元数量。结果缺血再灌注6h时，法舒地尔组MLK3磷酸化水平（1．13±0．03）显著低于生理盐水对照组（2．08±0．01，P=0．0003），但MLK3水平无显著差异（P=0．69）；缺血再灌注3d时，法舒地尔组JNK磷酸化水平（1．27±0．02）显著低于生理盐水对照组（2．09±0．01，P=0．0002），但JNK水平无显著差异（P=0．83）；缺血再灌注6h时，法舒地尔组胱冬酶-3表达水平（1．28±0．02）显著低于生理盐水对照组（2．10±0．01，P=0．0006）；缺血再灌注5d时，法舒地尔组海马CAl区存活锥体细胞数量为（82．8±3．2）个，显著多于生理盐水对照组的（11．8±1．6）个（P〈0．05）。结论法舒地尔能显著抑制缺血再灌注诱导的MLK3、JNK磷酸化水平以及胱冬酶-3蛋白表达，进而减轻海马CAl区神经元损伤。 Objective To investigate the effect of Rho kinase inhibitor fasudil on mixed lineage kinase 3 （MLK3）, c-Jun NH2-terminal kinase （JNK） phosphorylation, caspase-3 expression, and neuronal injury in hippocarnpal CA1 region follwong cerebral ischemic rep erfusion in rats. Methods A total of 72 Sprague-Dawley rats were randomly divided into sham operation, ischemia-reperfusion, normal saline, and fasudil groups. A global cerebral ischemic model was prepared by four-vessel ligation. The levels of MLK3 and JNK phosphorylation, and caspase-3 expression were detected by Western blot analysis. Cresyl violet staining was used to detect the numbers of survival neurons in hippocampal CA1 region. Results When 6 hours after ischemia-reperfusion, the level of MLK3 phosphorylation in the fasudil group （1.13 ± 0. 03） was sigaificantly lower than that in the normal saline group （2. 08 - 0. 01,P = 0. 000 3）, while the levels of MLK3 was no significant difference. When 3 hours after ischemia-reperfusion, the level of JNK phosphorylation in the fasudil group （1.27 - 0. 02） was significantly lower than that in the normal saline group （2. 09 - 0. 01, P = 0. 000 2）, while the levels of JNK was no signaificant difference. When 6 hours after ischemia-reperfusion, the expression level of caspase-3 in the fasudil group （1.28 ± 0. 02） was significantly lower than that in the normal saline group （2. 10±0. 01 ,P=0. 000 6）. When 5 days after ischemia-reperfusion, the pyramidal cells in hippocampal CA1 region almost completely disappeared in the ischemia-reperfusion group, and only a few cells left （9. 8 ± 2. 1）. The numbers of survival pyramidal cell （8. 28 ±3.2） in hippocampal CA1 region in the fasudil group was significantly more than that in the normal saline group （11.8 ± 1.6, P 〈 0. 05）. Conclusions Fasudil may significantly inhibit the ischemia-reperfusion-induced phosphorylation of MLK3 and JNK, as well as the expression of caspase-3, and thus reduce neuronal injury in hippocampal CA1 region.

作者魏秀娥张清秀荣良群刘春风

机构地区苏州大学附属第二医院神经内科徐州医学院第二附属医院神经内科

出处《国际脑血管病杂志》北大核心 2011年第1期69-74,共6页 International Journal of Cerebrovascular Diseases

关键词脑缺血法舒地尔神经保护药 p-相关激酶 MAP激酶激酶激酶类 JNK促有丝分裂激活蛋白激酶胱冬酶3 海马大鼠 Brain ischemia Fasudil Neuroprotective agents rho-associated kinases MAP kinase kinase kinases JNK mitogen-activated protein kinases Caspase 3 Hippocampus Rats

分类号 R74 [医药卫生—神经病学与精神病学] R9 [医药卫生—药学]

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